INTRODUCTION: Centrifuge-induced artificial gravity (AG) with exercise is a promising comprehensive countermeasure against the physiological de-conditioning that results from exposure to weightlessness. However, body movements onboard a rotating centrifuge are affected by both the gravity gradient and Coriolis accelerations. The effect of centrifugation on squat exercise biomechanics was investigated, and differences between AG and upright squat biomechanics were quantified.METHODS: There were 28 subjects (16 male) who participated in two separate experiments. Knee position, foot reaction forces, and motion sickness were recorded during the squats in a 1-G field while standing upright and while supine on a horizontally rotating 2 m radius centrifuge at 0, 23, or 30 rpm.RESULTS: No participants terminated the experiment due to motion sickness symptoms. Total mediolateral knee deflection increased by 1.0 to 2.0 cm during centrifugation, and did not result in any injuries. There was no evidence of an increased mediolateral knee travel "after-effect" during postrotation supine squats. Peak foot reaction forces increased with rotation rate up to approximately 200% bodyweight (iRED on ISS provides approximately 210% bodyweight resistance). The ratio of left-to-right foot force throughout the squat cycle on the centrifuge was nonconstant and approximately sinusoidal. Total foot reaction force versus knee flexion-extension angles differed between upright and AG squats due to centripetal acceleration on the centrifuge.DISCUSSION: A brief exercise protocol during centrifugation can be safely completed without significant after-effects in mediolateral knee position or motion sickness. Several recommendations are made for the design of future centrifuge-based exercise protocols for in-space applications.